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Amplification of SPPS150 and Salmonella typhi DNA with a high throughput oscillating flow polymerase chain reaction device
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10.1063/1.3422524
/content/aip/journal/bmf/4/2/10.1063/1.3422524
http://aip.metastore.ingenta.com/content/aip/journal/bmf/4/2/10.1063/1.3422524

Figures

Image of FIG. 1.
FIG. 1.

Schematic of oscillating flow PCR (not to scale).

Image of FIG. 2.
FIG. 2.

(a) Fabricated glass chip with nanoports attached to the access holes. (b) and (c) SEM of the microchannel bends.

Image of FIG. 3.
FIG. 3.

Schematic of the temperature control system.

Image of FIG. 4.
FIG. 4.

PCR chip system setup. (1) PCR glass chip, (2) syringe pump, (3) glass syringe, (4) fluidic connections, (5) temperature control system, and (6) heater housing with temperature sensors.

Image of FIG. 5.
FIG. 5.

Agarose gel electrophorogram of amplified 150 bp PCR product for cycle times of (a) 52.5, (b) 37.5, (c) 22.5, and (d) 29.5 min for 30 cycles (lane M: 100 bp ladder; lane C: control; lane 1: chip).

Image of FIG. 6.
FIG. 6.

Agarose gel electrophorogram of the amplified 1238 bp PCR product for cycle times of (a) 39.5 and (b) 55 min for 30 cycles (lane M: 1 kbp ladder; C: control; 1: chip).

Image of FIG. 7.
FIG. 7.

Gel electrophoresis results of DNA adsorption experiment; (a) 150 bp DNA templates (lane M: 100 bp ladder; lane C: control; lanes 1–4: chip) and (b) 1238 bp DNA templates (lane M: 1 kbp ladder; lane C: control; lanes 1–4: chip).

Image of FIG. 8.
FIG. 8.

Taq polymerase adsorption slab gel electrophoresis results. (a) 150 bp (lane M: 100 bp ladder; C: control; 1, 2, 3, 4: chip) and (b) 1238 bp (lane M: 1 kbp ladder; C: control; 1, 2, 3, 4: chip).

Image of FIG. 9.
FIG. 9.

Sample evaporation rate inside the PCR chip.

Image of FIG. 10.
FIG. 10.

(a) Formation of bubbles inside the microchannels at various temperatures without oil plug. (b) Absence of bubbles inside the microchannel even at high temperatures with the addition of oil plug.

Tables

Generic image for table
Table I.

Composition of different PCR mixtures.

Generic image for table
Table II.

Standard PCR temperature and cycle setup using conventional thermal cycler.

Generic image for table
Table III.

Experimental schemes for process optimization.

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/content/aip/journal/bmf/4/2/10.1063/1.3422524
2010-05-03
2014-04-23
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752b84549af89a08dbdd7fdb8b9568b5 journal.articlezxybnytfddd
Scitation: Amplification of SPPS150 and Salmonella typhi DNA with a high throughput oscillating flow polymerase chain reaction device
http://aip.metastore.ingenta.com/content/aip/journal/bmf/4/2/10.1063/1.3422524
10.1063/1.3422524
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